Kinetic study of enzymatic α-galactoside hydrolysis in cowpea seeds.

The endogenous alpha-galactosidase activity of cowpea seeds was characterized and modelled assuming Michaelian behavior. The aim is to use the resulting knowledge to optimize alpha-galactoside degradation during the soaking-cooking process. In this study, the alpha-galactosidase enzyme from Wankoun cowpea was extracted and its enzymatic activity was analyzed as a function of temperature, pH and the presence of some inhibitors. Enzymatic activity was optimal around 35 °C and a pH of 5.8. Activation and inactivation energy was evaluated at 50 ± 3 and 103 ± 9 kJ.mol-1 , respectively. The strongest inhibitor was galactose with an inhibition constant KI of 0.28 ± 0.03 mM. Incubation of the enzyme extract with alpha-galactosides revealed a 10-h lag phase in the early stages that could be due to low pH, the action of inhibitors including galactose and the biosynthesis of alpha-galactosides. After the lag phase, the degradation of each alpha-galactoside occurred without the appearance of any intermediary product. The degradation of alpha-galactosides was observed with a Km of 1.7 ± 0.3 mM for raffinose; 3.6 ± 0.6 mM for stachyose and 15.9 ± 0.1 mM for verbascose. A long soaking step around 35 °C is suggested to maximize the alpha-galactosides enzymatic degradation.